Fucose

This antibody specifically cross-reacts against fucose residues bound to the protein N-glycans in alpha 1,3. This residue is characterisitc of the plant protein N-glycans and is absent in protein N-glycans from animals. This residue is added in the Golgi apparatus.

Immunogen

core fucose residues bound to the N-glycan in alpha 1,3

Host

Rabbit

Clonality

Polyclonal

Clone

Purity

Affinity purified serum in PBS, pH 7.4

Format

Lyophilized in PBS pH 7.4

Quantity

50 µg

Reconstitution

For reconstitution add 50 µl of sterile water.

Storage

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Alpha (1,3) fucose is present not only in plants but also in some invertebrates (such as nematodes, bees, etc.) . However, cross-reaction with glycoproteins from these organisms is weaker than the one observed in plants. This sugar residue does not exist in mammals, in their endogenous glycoproteins.

application information

Recommended dilution

0.5 µg/ml (ELISA), 1 : 40 (IL), 1 ug/10 ml (WB)

Expected | apparent MW

10 - 100 for various glycoproteins

Confirmed reactivity

Higher plants

Predicted reactivity

Higher plants

Not reactive in

No confirmed exceptions from predicted reactivity are currently known.

The Anti-fucose antibodies exhibit a positive response with PLA2 and Avidin and a negative one with asialofetuin. This suggests this antibody recognizes only α1,3 fucose epitopes.

For each sample, 1µg of purified protein was denatured with leammli sample buffer at 100°C for 10 minutes loaded on a Bis-Tris gel 4-12% and blotted 1h15 to nitrocellulose membrane using semi-dry transfer. Blots were saturated in TBST over night with agitation. Blot was incubated for 2 hours with primary antibody at a dilution of 1:5000 in TBST at RT with agitation. The antibody solution was discarded and after quick whashes with TBST, the blot was washed 3X15 min in TBST. Blot was incubated in secondary antibody (Goat anti Rabbit IgG, HRP conjugated AS09 602 lot 1707 from AGRISERA) diluted to 1:50 000 in TBST for 1 hour at RT with agitation. The blot was washed as above, plus 1 wash in TBS for 10 min at RT with agitation. The blot was developed for 5 min with the kit ECL West Pico Plus from Thermofisher. Exposure time was 5 seconds.